Hydrogel-based vascular lineage cell growth media and uses thereof

Abstract

A medium for growing vascular lineage cells is described. The vascular lineage cell growth medium includes an oligosaccharide-based hydrogel and a growth factor that promotes vascularization by vascular lineage cells.

We claim: 
     
       1. A method of preparing a vascular lineage cell growth medium comprising
 chemically bonding an adhesion promoter to an oligosaccharide, 
 combining the chemically bonded adhesion promoter and oligosaccharide with at least one growth factor, 
 adding a crosslinking peptide moiety, 
 curing the combination to form a hydrogel, wherein the hydrogel has a Young's modulus between about 10 Pa and about 500 Pa; and 
 wherein the growth factor is an amount sufficient to promote vasculogenesis and tube formation, and sufficient to stimulate MMP production in vascular lineage cells. 
 
     
     
       2. The method of  claim 1 , wherein the oligosaccharide is hyaluronic acid that is from about 20% to about 90% acrylate modified and is present in an amount of about 1 wt % to about 4 wt %. 
     
     
       3. The method of  claim 1 , wherein the adhesion promoter is present at a concentration of from about 0.1 mM to about 20 mM. 
     
     
       4. The method of  claim 1 , wherein the crosslinking peptide moiety is present at a concentration of from about 1 mM to about 8 mM. 
     
     
       5. The method of  claim 1 , wherein the growth factor is present at a concentration of from about 25 ng/ml to about 200 ng/ml. 
     
     
       6. The method of  claim 1 , wherein the hydrogel allows formation of capillary-like structures (CLSs) with extended vacuoles and open lumens from vascular lineage cells. 
     
     
       7. The method of  claim 1 , wherein the hydrogel and crosslinking peptide moiety are cleavable by an enzyme. 
     
     
       8. The method of  claim 7 , wherein the enzyme is a matrix metalloproteinase. 
     
     
       9. The method of  claim 8 , wherein the matrix metalloproteinase is at least one of MMP-1, MMP-2 or MMP-10. 
     
     
       10. The method of  claim 1 , wherein the crosslinking moiety peptide comprises the sequence SEQ ID NO: 1. 
     
     
       11. The method of  claim 1 , wherein the adhesion promoter promotes vascular lineage cell adhesion to the hydrogel and promotes vacuole and lumen formation in vascular lineage cells. 
     
     
       12. The method of  claim 1 , wherein the hydrogel has a Young's modulus between about 10 Pa and about 250 Pa. 
     
     
       13. The method of  claim 1 , wherein the oligosaccharide is modified with an acryl group. 
     
     
       14. The method of  claim 1 , wherein the oligosaccharide is acrylated hyaluronic acid. 
     
     
       15. The method of  claim 1 , wherein the growth factor is selected from the group consisting of VEGF, TNFα, SDF1-α, bFGF, angiopoetin-1, PDGF, TGF-β, PIGF or combinations thereof. 
     
     
       16. The method of  claim 1 , wherein the adhesion promoter is a peptide that includes an RGD sequence and a reactive functional group that is capable of bonding the adhesion promoter to the oligosaccharide. 
     
     
       17. A vascular lineage cell growth medium prepared by the method of  claim 16 , wherein the oligosaccharide is acrylated hyaluronic acid. 
     
     
       18. A method of inducing vascular cell morphogenesis in vivo comprising contacting a subject with a vascular construct comprising the medium of  claim 17  and vascular lineage cells. 
     
     
       19. The method of  claim 18 , wherein the crosslinking moiety peptide comprises the sequence SEQ ID NO: 1. 
     
     
       20. The method of  claim 18 , wherein the growth factor is selected from the group consisting of VEGF, TNFα, SDF1-α, bFGF, angiopoetin-1, PDGF, TGF-β, PIGF or combinations thereof. 
     
     
       21. The method of  claim 1 , wherein the oligosaccharide is modified with a thiol group.